At pH 6.0 DTT does not exist in its thiolate form. As thiolate form of DTT initiates the reduction process, can it still perform its function at pH 6.0? Or are there any other alternative reducing agents that could be used at pH 6.0, phosphate buffer?
Most references say DTT doesn't function at acidic pH, but it has a weak reducing property till around 5.5 .. It is better to use pH 7, although I've seen quite a few papers on enzyme characterization where DTT has been used at pH
Thanks Avinash. Can I use TCEP? Would it react with phospahte buffer?
TCEP is a little expensive, but it is more stable over wide pH range and more effective reducing agent compared to DTT. The Pierce catalogue says TCEP is not stable in PBS at neutral pH, but degradation minimal at pH
Just an additional data : if you begin with TCEP, do not add DTT at any part of your experiment. You might have some "strange" reaction.
TCEP and/or 2-mercaptoethanol may be used. We use these reducing agents to extract keratin from hair by breaking the disulphide bonds. use it in combination with a small amount of thiourea.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Molecular Biological Techniques